ADCs have evolved since the initial approval of MYLOTARG® in 2000, with respect to both conjugation, linker and toxin chemistries as well as processing. With the current availability of four commercially approved drugs and approximately 80 programs in various clinical trials, there has been a significant interest in simplifying the complex supply chain to make manufacturing efficient. With approximately 80% of the programs outsourced to Contract Development and Manufacturing Organizations (CDMOs), a transparent and integrated supply chain is critical for the success of ADC projects. MilliporeSigma has been involved with the development and manufacturing of ADCs since 2008. This webinar will discuss how chemistry and manufacturing have evolved over the past 10 years globally and how MilliporeSigma, as a CDMO has adapted to these changes to meet customers needs. In this webinar, you will learn: ● How ADC manufacturing has evolved over the past decade ● The challenges that this evolution implies ● Why an integrated supply chain has become increasingly important for the success of ADC projects

1. Merck KGaA
Darmstadt, Germany
Jyothi Swamy and Lisa McDermott
April 5, 2018
THE 10-YEAR
JOURNEY EVOLVING
TO A COMMERCIAL
ADC MANUFACTURER

2. 2
The life science business of
Merck KGaA, Darmstadt, Germany
operates as MilliporeSigma
in the U.S. and Canada.
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

3. Evolution of ADCs
Outline
Introduction to ADCs
A Few Facts about ADCs
Chemistry – Then, Now and Next
Changes since 2000
Summary
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 20183

4.  Antibody
 Payload
 Linker
Specific for a tumor-associated
antigen that has restricted
expression on normal cells.
Designed to kill target cells
when internalized and
released.
Attaches the cytotoxic agent to the
antibody. New linker systems are
designed to be stable in circulation
and release the cytotoxic agent
inside targeted cells.
Introduction to ADCs
Elements of an ADC
Conjugation
is the step of linking
the cytotoxic agent to
the antibody.
4 Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

5. 2018 &
beyond
2017
Global History
ADC Approvals
2011 2013
2000/
2017*
MylotargTM
 Pfizer
Adcetris®
 Seattle
Genetics/Takeda
Kadcyla®
 Genentech
(Hoffmann
LaRoche)
BesponsaTM
 Pfizer
Which one is
next?**
5
**Experts1 in the
field predict that the
following to gain
approval in the near
future
a) Sacituzumab
govitecan
b) Rovalpituzumab
tesirine
c) Glembatumumab
Vedotin
d) Mirvetuximab
soravtansine
*Initially
approved in
2000 and
withdrawn from
US & EU markets
in 2010. Based
on the new
clinical data, FDA
re-approved in
September 2017
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018
1Beacon reports & alerts and P. Drake & D Rabuka, Bioprocess Online, March 6, 2018

6. A Few Facts About ADCs
1Beacon Reports and alerts, World ADC Conferences
*YTD
6
ADC Pipeline – Globally1
Total Number of Active Clinical Programs
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

7. • 4 commercially approved products
• ~75 programs in various clinical
Phases
• >100 pre-clinical programs
• 70 to 80% of ADC projects are
outsourced for various reasons
• No ADC specific guidelines from
FDA or EMEA
• More clinical trials with combination
therapy expected (>200 trials
registered)
• ADCs beyond Oncology expected to
gain prominence:
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 20187
- Anti-infection (Antibody antibiotic conjugate)
- Cardiovascular Diseases
- Imaging & Diagnostic Agents

8. 8
Poll Question

9. Testing Flow
Material Flow
Supply Chain Complexity
9
mAb
Production
Linker
Payload
Production
Custom Raw
Materials
(Linker &
Payload)
Conjugation
Bioassay
Testing
Stability
Testing
Cold
Storage
Label,
Pack &
Clinical Use
Formulation
& Filling
 Anywhere from 5-10 CMO’s involved
 Multi-geographical footprint & logistics

10. 10
Available Global ADC Technologies
Cytotoxic Agent• Humanized IgG
• Site Specific Engineering
• Bi-Specific mAbs
• Fab
• Engineered NNAA
• mAb Fragments
• Cleavable
• Enzymatic (protease)
• Acid Labile (hydrazone)
• Non-Cleavable
• PEG/Cyclohexyl
• Hindered Disulfide
• Microtubule Disruption
• Maytansines
• Auristatins
• Auristatin anlogs
• Tubulysins
• DNA/RNA Damaging Agents
• Calicheamicin
• Doxorubicin
• PBDs
• Duocarmycins
• Indolinobenzodiaepines
• Topoisomerase inhibitor
• SN-38
• Amanitin
Antibody
Linker
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

11. 11
ADC History
Experience with 43 different constructs and >300 batches in Development
With Significant emphasis on Chromatographic Purifications
Random cysteine or lysine conjugation technology
Site directed conjugation via engineered mAbs, enzyme catalyzed or
non-natural amino acids
Various payloads and Linker chemistries
Advances in Analytical Techniques and Regulatory Experience
First Full Single Use GMP Manufacture in 2017
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

12. 12
GMP ADC Production
# of Batches
(g input mAb)
30 - 100 20
101 - 500 54
>500 22
GMP batches 96
ADC Constructs > 25
800 sq.ft Clinical Manufacturing Facility Suite
10,000 sq.ft Commercial Manufacturing Facility Suite
Dedicated QA, QC and Regulatory Resources
Safebridge Certified Site
Our ADC Expertise: Summary
ADC Manufacturing Expertise
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

13. Lysine Addition
 Heterogeneous Product
 Insufficient Analytical Tools
Random Cysteine
 Loss of Payload due to Retro Michael Addition
 High DAR species
Stability
PK/PD Profile
Processing Issues
Common Issues
 Purification limited to TFF
 Technology Built onto mAb platforms vs optimized for conjugation
Conjugation Chemistry
Then
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 201813

14. mAb Advancement
• Engineered Cysteine, Non-Natural Amino Acid, Enzyme Mediated
• Site Specific conjugation
• First Site Specific Conjugate in Clinic -2013 (SGN-CD33A)
• Controlled DAR 2 and 4 Species Distribution
 Stability, Toxicity, Clearance differences
• Tighter distribution
• Aggregation control
• SAR Drug Design
Conjugation Chemistry
Now and Beyond
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 201814

15. Drug Advancement
• Payload MOA
• More Potent Cytotoxic Payloads
• First PBD Conjugate in Clinic - 2013
Linker Advancement
• Modulation of ADC hydrophobicity with hydrophilic Linkers
• Optimized on an individual basis
General Advancements
• Enhanced Analytical tools
• Large Scale Chromatography
• Technology Customized to ADCs
Conjugation Chemistry
Now and Beyond
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 201815

16. 16
Poll Question

17. Antibody
Humanized IgG’s
• Full Length mAbs
• IgG1 >> IgG2 / IgG4
• Random Conjugates
Site Specific Conjugates
• Engineered Cysteine
• Non-Natural Amino Acids
• Sequence Tags
- Enzyme Mediated
- SMARTagTM Platform
Antibody Analogs
• Bi-Specific mAbs
• mAb Fragments
Payloads
First Generation
• Calicheamicin
• SN38
• Duocarmycin
• Doxorubicin
Second Generation
• Auristatins
• Maytansines
Third Generation
• PBD’s
First Generation
• Monovalent
• Non-Cleavable
• Acid Labile
Second Generation
• Monovalent
• Cleavable – Peptidase
• Cleavable - Disulfide
Third Generation
• Fleximer Platform
• Polyvalent
• Bio-Degradable
Linkers
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 201817
ADC Building Blocks
Evolution of Technology / Experience Footprint

18. GMP Manufacturing Scale Staffing Challenges
100-500g / batch Experience or “Fit”
Limited Commercial Experience
Clinical Supplies
Limited Regulatory History
Limited High Potent Capabilities
Drug Linker Manufacturers
Biological Manufactures
Modeled after mAb backbone
Analytics
Process Unit Operations
Manufacturing
Then
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018
18

19. Larger scale – 1-3Kg/batch
Expanded due to multiple trials and combinations
Better understanding of pre-clinical models
Commercial Experience
Regulatory Expectations
ie: Species Distribution, Aggregate Control
Speed from FIH to Commercial (Breakthrough)
CMC on critical path vs Clinical Studies
Single-use Technology
Complete SU Process Train
Disposable UV Flow Paths, on line process monitoring
Reduced Cleaning Studies, Reduced Costs
19
Manufacturing
Now and Beyond
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

20. Process and Analytical Technology (PAT)
On line and at line for process understanding
Availability of Platform Equipment
Semi –Custom Skids for larger scale
TFF, Chromatography
Distribution Controlled by Chromatography
Increase Number of Constructs have Chromatography
CHT, SCX, HIC, Mixed Mode
20
Manufacturing
Now and Beyond
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

21. Single-Use ADC Advancements
Upcoming Webinar April 19th:
Case Study of Full Single Use
Process in GMP

22. ADCs: Unique Regulatory Space
Impurities
Pre and Post
Conjugation
Free Drug
and Related
Species
API-like
Control of mAb,
Linker and
Drug
Loading
and
Distribution
Free vs
Conjugated
mAb
Unique
Stability
Profiles
Potency
Pre and Post
Conjugation
Controls of
Size and
Distribution
Adapting Analytical Control Strategy to Project Pace
Validation per Phase of Development vs Accelerated Path

23.  Conjugation Chemistry moving toward Site Specific Constructs
 Increased toxicity of Payloads
 Higher diversity of Linkers and Impact on Conjugation, Drugability and Polydispersity
 Increased Manufacturing Scale
 Regulatory and Commercial Experience
 Moving toward Single Use Across all Unit Operations
 Advancement of Analytical Tools and PAT
 Implementation of Chromatographic Purification for Distribution Control
 Acceleration between FIH and Commercial
23
Summary of Manufacturing Changes
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

24. 24
• Next Changes in ADCs and the impact on Manufacturing
• Increased need for advanced analytical tools and PAT
• Implementation of Single Use Reactors
• Advances in Chromatographic Purifications and Equipment
What’s Next?
Webinar: The 10-year Journey evolving to a commercial ADC Manufacturer | April 5, 2018

25. Jyothi Swamy
jyothi.swamy@sial.com
The Vibrant M is a trademark of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the
property of their respective owners. Detailed information on trademarks is available via publicly accessible
resources. © 2018 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.
Lisa McDermott
lisa.mcdermott@sial.com